基底细胞是气道上皮中的成体干细胞，可再生分化的细胞群，包括负责粘液纤毛清除的粘液分泌细胞和纤毛细胞。人类基底细胞可以在体外增殖并产生分化的上皮。然而，气道上皮分化的研究主要依赖于免疫组织化学或基于免疫荧光的染色方法，这意味着缺乏动态方法，并且定量数据有限。在这里，我们使用慢病毒报告基因方法用生物发光报告基因构建物转导原代人类基底细胞，以纵向监测气道上皮分化。我们生成了由 TP63、MUC5AC 和 FOXJ1 基因的启动子序列驱动的三个构建体，分别定量评估基底细胞、粘液分泌细胞和纤毛细胞丰度。我们通过跟踪气液界面和类器官（“支气管球”）培养物中基底细胞的分化来验证这些构建体。转导细胞也对白细胞介素 13（IL-13；增加粘液分泌分化和粘液产生）和 IL-6（增加纤毛细胞分化）的刺激做出适当反应。这些构建体代表了一种用于监测原代上皮细胞和/或诱导多能干细胞（iPSC）细胞培养物中气道上皮细胞分化的新工具。

Basal cells are adult stem cells in the airway epithelium and regenerate differentiated cell populations, including the mucosecretory and ciliated cells that enact mucociliary clearance. Human basal cells can proliferate and produce differentiated epithelium in vitro. However, studies of airway epithelial differentiation mostly rely on immunohistochemical or immunofluorescence-based staining approaches, meaning that a dynamic approach is lacking, and quantitative data is limited. Here, we use a lentiviral reporter gene approach to transduce primary human basal cells with bioluminescence reporter constructs to monitor airway epithelial differentiation longitudinally. We generated three constructs driven by promoter sequences from the TP63, MUC5AC and FOXJ1 genes to quantitatively assess basal cell, mucosecretory cell and ciliated cell abundance, respectively. We validated these constructs by tracking differentiation of basal cells in air-liquid interface and organoid ('bronchosphere') cultures. Transduced cells also responded appropriately to stimulation with interleukin 13 (IL-13; to increase mucosecretory differentiation and mucus production) and IL-6 (to increase ciliated cell differentiation). These constructs represent a new tool for monitoring airway epithelial cell differentiation in primary epithelial and/or induced pluripotent stem cell (iPSC) cell cultures.